Effect of natural protease inhibitors and a chemoattractant on tumor cell invasion in vitro.
Effect of natural protease inhibitors and a chemoattractant on tumor cell invasion in vitro.
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天然蛋白酶抑制剂和趋化剂对体外肿瘤细胞侵袭的影响。
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发表时间：
1982
期刊：
影响因子：
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通讯作者：
R. G. Russo
中科院分区：
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作者：
U. Thorgeirsson;L. Liotta;T. Kalebic;I. Margulies;K. Thomas;M. Rios;R. G. Russo
利用一种新的肿瘤细胞穿透天然结缔组织的体外定量测定法，研究了天然蛋白酶抑制剂和化学引诱剂对肿瘤细胞侵袭的影响。人羊膜剥除上皮后，由连续的基底膜（BM）和致密的无血管胶原基质组成。将已知在体内具有高度侵袭性的M5076网状肉瘤细胞置于羊膜结缔组织的BM侧。用Millipore过滤器在基质侧收集穿透结缔组织屏障全层的肿瘤细胞。与无血清培养基中自发穿透的肿瘤细胞数量相比，最佳浓度为10（-7）M的N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸（FMLP）在20小时后刺激了高达600%的肿瘤细胞穿透进入结缔组织。天然蛋白酶抑制剂阻断了FMLP刺激的和自发的侵袭。一种含有丝氨酸蛋白酶和金属蛋白酶抑制剂的牛软骨提取物导致侵袭减少500%。此外，纯化的胶原酶（金属蛋白酶）抑制剂可对侵袭产生500%的抑制作用。相反，大豆胰蛋白酶抑制剂和牛血清白蛋白没有显着改变侵袭率。蛋白酶抑制剂是无毒的，并且不会减少肿瘤细胞增殖、与羊膜的附着以及肿瘤细胞通过Nuclepore过滤器的迁移速率。这些数据支持这样的假设，即胶原溶解性金属蛋白酶在肿瘤细胞侵袭天然结缔组织中起着必要的作用。
The effect of natural protease inhibitors and a chemoattractant on tumor cell invasion were studied with the use of a new in vitro quantitative assay of tumor cell penetration of native connective tissue. Human amnion membrane denuded of its epithelium is composed of a continuous basement membrane (BM) attached to a dense avascular collagenous stroma. M5076 reticulum sarcoma cells, known to be highly invasive in vivo, were placed on the BM side of the amnion connective tissue. Tumor cells penetrating the full thickness of the connective tissue barrier were collected on the stromal side with a Millipore filter. N-Formylmethionyl-leucyl-phenylalanine (FMLP) at an optimal concentration of 10(-7) M stimulated the penetration of up to 600% more tumor cells into the connective tissue after 20 hours in comparison to the number of tumor cells spontaneously penetrating in serum-free media. Natural protease inhibitors blocked both FMLP-stimulated and spontaneous invasion. A bovine cartilage extract containing inhibitors of both serine proteinases and metalloproteinases caused a 500% decrease in invasion. Furthermore, a 500% inhibition of invasion was produced by a purified collagenase (metalloproteinase) inhibitor. In contrast, soybean trypsin inhibitor and bovine serum albumin did not significantly alter the invasion rate. The protease inhibitors were nontoxic and did not reduce tumor cell proliferation, attachment to the amnion, and the rate of tumor cell migration through Nuclepore filters. These data support the hypothesis that collagenolytic metalloproteinases play a necessary role in tumor cell invasion of native connective tissue.